Microorganism resistance to synthetic antibiotics is an increasing public health challenge; therefore, Researchers are in strong interest to develop new antimicrobial agents from biological sources. It is well studied that mushrooms are being used since ancient time as a means of food and medicine. Mushrooms are potent source of various bioactive compounds. Antimicrobial activities have been discovered in a wide range of secondary metabolites, including tannins, terpenoids, alkaloids, and flavonoids. These natural compounds are of concern as a source of safer and more effective antibacterial agents than those produced synthetically. Different extracts of mushroom has a significant amount of bioactive compounds which may contribute to antibacterial properties. In present study we have evaluated the different phytochemicals in different extracts of mushroom “Clitocybe phaeophthalma”. The collected variety of mushroom were washed carefully, air dried and powdered for further downstream process. The prepared powder from mushroom was subjected to isolate different extracts using Soxhlet extractor, using different solvent system. Different crude drug extracts of the sample were used for biochemical analysis to detect the phytochemical constituents in the fungi. The results of the present study of showed the presence of different phyto-constituents such as flavonoids, terpenoids, and alkaloids etc. The Qualitative analysis has confirmed the presence of the antimicrobial phytochemicals in the Clitocybe phaeophthalma.